JP2024023213A - JAK1 pathway inhibitors to treat cytokine-related disorders - Google Patents

Abstract

The present invention provides a medicament for treating CAR-T cell-related encephalopathy syndrome.
[Solution] A pharmaceutical comprising a JAK1 pathway inhibitor, wherein the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3 -[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof, It is used as a medicine. Preferably, said medicament is administered in combination with tocilizumab and/or corticosteroids.
[Selection diagram] None

Description

The present disclosure relates to JAK1 pathway inhibitors and their use in treating cytokine-related diseases or disorders.

Cytokine-related diseases or disorders are characterized by excessive activation of immunity and include cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS) and CAR-T cell-associated encephalopathy syndrome. (CRES).

Cytokine release syndrome (CRS) is a direct result of overproduction of inflammatory cytokines caused by supraphysiological levels of immune activation, resulting in fever, nausea, fatigue, myalgia, malaise, low blood pressure, It manifests as a series of clinical symptoms such as hypoxia, capillary leakage, thereby leading to potential multisystem toxicity.

CRS is an unwanted side effect of immunotherapy for serious disease conditions such as cancer. Immunotherapy that can cause CRS includes the administration of monoclonal antibodies (mAbs) and, more recently, adoptive T cell therapy for cancer. Lee et al. Blood. 2014, 124(2): 188-195. For example, chimeric antigen receptor (CAR) T-cell therapy uses engineered T cells to target cancer and treats certain forms of refractory non-Hodgkin's lymphoma and childhood relapsed lymphoblastic leukemia (ALL). ) has already been approved by the FDA for use in

The cytokine profile involved in CRS includes two major cellular sources. It contains T lymphocyte-derived cytokines, including interferon gamma (IFN)-γ, IL-2, IL-6, soluble IL-6 receptor (IL-6R) and granulocyte-macrophage colony-stimulating factor (GM-CSF); Cytokines such as IL-1β, IL-6, IL-12, IL-18 and tumor necrosis factor (TNF)-α are mainly secreted by monocytes and/or macrophages. Xu XJ, Tang YM. Cancer Lett. 2014;343:172-8. Zhang Y. , et al. Sci China Life Sci. 2016;59:379-85. Brentjens R. , et al. Mol Ther. 2010;18:666-8.

Modulating the exaggerated cytokine responses that lead to CRS has the potential to provide important clinical benefits. For example, tocilizumab, an antibody against the IL-6 receptor (IL-6R), reduces the incidence of severe CRS and has been approved by the FDA for use in CRS. However, the mechanism of action of tocilizumab is limited to anti-IL-6R.

Hemophagocytic lymphohistiocytosis (HLH) is another syndrome of excessive or uncontrolled immune activation that primarily affects infants from birth to 18 months of age, but may also affect adults. HLH can be primary (familial) or secondary, meaning that it develops in the context of other infectious, malignant, rheumatic or metabolic conditions. Symptoms of HLH include cytopenias, hepatosplenomegaly, and fever. Schram, A. and Berliner, N. Blood. 2005.125(19), 2908-2914.

Macrophage activation syndrome (MAS) presents clinically in a similar manner to HLH (further considered secondary or acquired HLH), due to increased inflammation associated with infection, rheumatic disease or malignancy. develop. Borgia, R. E. et al. Arthritis Rheumatol. , 2018, doi:10.1002/art. 40417 (before publication). Although MAS was first described as associated with juvenile idiopathic arthritis, it is increasingly recognized as a complication of other diseases such as childhood-onset systemic lupus erythematosus (cSLE). Shimizu M. , et al. Clin Immunol. 2013 Feb; 146(2):73-6. The development of MAS is characterized by a substantial increase in a number of pro-inflammatory cytokines, ie, a cytokine storm. Borgia, R. E. et al. Arthritis Rheumatol. , 2018, doi:10.1002/art. 40417 (before publication). MAS is a serious condition with a high mortality rate, typically 8-22% for pediatric autoimmune diseases and 10-22% for MAS complicated by cSLE. Borgia, R. E. et al. Arthritis Rheumatol. , 2018, doi:10.1002/art. 40417 (before publication).

CAR-T cell-related encephalopathy syndrome (CRES) is the second most common adverse event after CRS associated with CAR-T cell therapy. CRES is usually characterized by a state of toxic encephalopathy with symptoms of confusion and delirium, as well as occasional seizures and cerebral edema. Symptoms of CRES are biphasic and can occur within the first 5 days and/or 3-4 weeks after cellular immunotherapy. The pathophysiological mechanism is believed to include passive diffusion of cytokines into the brain of patients treated with CAR-T cell therapy. Reducing or eliminating this mechanism may be beneficial for such patients. Neelapu, et al. Nat Rev Clin Oncol. 2018, 15(1) 47-62.

Therefore, there is a need to develop new therapeutics to treat cytokine-related diseases or disorders. The present application addresses this and other needs.

Figure 3 shows a dose-dependent inhibition of IL-6 concentrations upon administration of Compound 1 into the blood compartment during anti-CD3 antibody-induced cytokine release syndrome (see Example B). Figure 2 shows dose-dependent inhibition of T cell-derived cytokines (ie, IL-6) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). Inhibition of IL-6 is shown. Figure 3 shows dose-dependent inhibition of T cell-derived cytokines (i.e., IFNγ) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). Inhibition of IFNγ is shown. Figure 3 shows dose-dependent inhibition of T cell-derived cytokines (ie, GM-CSF) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). Inhibition of GM-CSF is shown. Figure 3 shows dose-dependent inhibition of monocyte- and/or macrophage-derived cytokines (ie, IL-12) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). Inhibition of IL-12 is shown. Figure 3 shows dose-dependent inhibition of monocyte- and/or macrophage-derived cytokines (ie, IL-1β) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). Inhibition of IL-1β is shown. Figure 3 shows dose-dependent inhibition of monocyte- and/or macrophage-derived cytokines (ie, IL-18) upon administration of Compound 1 during concanavalin A-induced cytokine release syndrome (see Example C). Showing inhibition of IL-18. The cytokine IL-5 is shown to have no effect on Compound 1 treatment during concanavalin A-induced cytokine release syndrome (see Example C).

Provided herein are methods of treating a cytokine-related disease or disorder in a subject in need of treatment, which methods include administering to the patient a therapeutically effective amount of a JAK1 pathway inhibitor or a pharmaceutically acceptable amount thereof. This includes administering salt.

Provided herein are JAK1 pathway inhibitors or pharmaceutically acceptable salts thereof for treating cytokine-related diseases or disorders in a subject in need of treatment.

Provided herein is the use of a JAK1 pathway inhibitor, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for use in treating a cytokine-related disease or disorder in a subject in need of treatment.

Detailed description of the invention

The present invention provides, inter alia, a method of treating a cytokine-related disease or disorder in a subject in need of treatment, which method comprises administering to the subject a therapeutically effective amount of a JAK1 pathway inhibitor or a pharmaceutically acceptable amount thereof. This includes administering suitable salts.

The methods described herein utilize JAK1 pathway inhibitors, particularly JAK1 selective inhibitors. JAK1 selective inhibitors are compounds that preferentially inhibit JAK1 activity over other Janus kinases. JAK1 plays a central role in many cytokine and growth factor signaling pathways that, when dysregulated, can lead to or contribute to disease states. For example, IL-6 levels are elevated in rheumatoid arthritis, and it has been suggested that IL-6 has deleterious effects in this disease (Fonesca, et al., Autoimmunity Reviews, 8:538-42, 2009 ). Because IL-6 signals at least in part through JAK1, IL-6 exists indirectly through inhibition of JAK1, providing potential clinical benefit (Guschin, et al. Embo J 14: 1421, 1995; Smolen, et al. Lancet 371:987, 2008). Furthermore, in some cancers, JAK1 is mutated, resulting in constitutive undesirable tumor cell proliferation and survival (Mulligan, Proc Natl Acad Sci USA. 106:9414-8, 2009; Flex, J Exp Med. 205:751-8, 2008). In other autoimmune diseases and cancers, elevated systemic levels of inflammatory cytokines that activate JAK1 may also contribute to the disease and/or associated symptoms. Therefore, patients with such diseases may benefit from inhibition of JAK1. Selective inhibitors of JAK1 may be beneficial while avoiding unnecessary and potentially undesired effects of inhibiting other JAK kinases.

JAK1 pathway inhibitors, specifically compound 1 (i.e., {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo [2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile (see Table 1) is a highly effective inhibitor of CRS-related inflammatory cytokines. (See, eg, Examples B and C and Figures 1, 2A-2C and 3A-3C). Surprisingly, this therapeutic property includes multiple pathogenic cytokines and is not limited to only the IL-6/IL-6R system (eg, tocilizumab is different). Efficacy is achieved by inhibiting cytokines derived from T cells and monocytes/macrophages that are clinically highly relevant to the pathogenesis of CRS. Furthermore, the data presented herein related to the JAK1 inhibitor Compound 1 indicate that therapeutic benefit is achieved without extensive cytokine immunosuppression (IL-5 levels remain unchanged). (Figure 4).

In some embodiments, the cytokine-related disease or disorder is cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell-associated encephalopathy syndrome (CRES). ).

In some embodiments, the cytokine-related disease or disorder is cytokine release syndrome (CRS).

In some embodiments, the cytokine-related disease or disorder is hemophagocytic lymphohistiocytosis (HLH).

In some embodiments, the cytokine-related disease or disorder is macrophage activation syndrome (MAS). In some embodiments, macrophage activation syndrome is associated with systemic juvenile idiopathic arthritis. In some embodiments, macrophage activation syndrome is associated with childhood systemic lupus erythematosus.

In some embodiments, the cytokine-related disease or disorder is CAR-T cell-related encephalopathy syndrome (CRES).

In some embodiments, the present application provides a method of treating cytokine release syndrome in a subject, the method comprising administering to the subject CAR-T cell therapy and a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof. including administering. In some embodiments, treating is ameliorating or inhibiting. In some embodiments, treatment is prevention.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered concurrently with CAR-T cell therapy.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered after administration of CAR-T cell therapy.

In some embodiments, the CAR-T cell therapy is axicabtagene ciloleucel.

In some embodiments, the CAR-T cell therapy is tisagenlecleucel.

In some embodiments, the subject has a B cell malignancy.

In some embodiments, the subject has diffuse large B-cell lymphoma (DLBCL), primary mediastinal large B-cell lymphoma, high-grade B-cell lymphoma, transformed follicular lymphoma, or acute lymphoblastoma. He has leukemia.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is selective for JAK1 over JAK2, JAK3 and TYK2 (ie, a JAK1 selective inhibitor). For example, a compound described herein, or a pharmaceutically acceptable salt thereof, preferentially inhibits JAK1 over one or more of JAK2, JAK3 and TYK2. In some embodiments, the compound preferentially inhibits JAK1 over JAK2 (eg, JAK2/JAK1 IC ₅₀ ratio >1). In some embodiments, the compound or salt is about 10-fold selective for JAK1 over JAK2. In some embodiments, the compound or salt is about 3-fold, about 5-fold, about 10-fold, about 15-fold, about 15-fold or about 20-fold more potent against _JAK1 than JAK2, as calculated by measuring the IC 50 at 1 mM ATP. Fold selective (see, eg, Example A).

In some embodiments, the JAK1 pathway inhibitor is a compound of Table 1 or a pharmaceutically acceptable salt thereof. The compounds in Table 1 are selective JAK1 inhibitors (more selective than JAK2, JAK3 and TYK2). The IC ₅₀ values obtained by the method of Example A at 1 mM ATP are shown in Table 1.

+ Average ≦10 nM (see Example A for assay conditions)
++ Average ≦100 nM (see Example A for assay conditions)
+++ Average ≦300 nM (see Example A for assay conditions)
Data for ^a enantiomer 1
Data for ^b enantiomer 2

In some embodiments, the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[ 2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof.

In some embodiments, the JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[ 2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate.

{1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)- The synthesis and preparation of 1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile and its adipate salts are described in US Patent Application No. 2011/0224190 (filed March 9, 2011), US Patent Application No. 2013/ No. 0060026 (filed September 6, 2012) and U.S. Patent Application No. 2014/0256941 (filed March 5, 2014), each of which is incorporated herein by reference in its entirety. .

In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl) azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof.

In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl) azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide phosphate.

4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro- The synthesis and preparation of N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide and its phosphate salts are described, for example, in US Patent Application No. 2014/0343030 (May 16, 2014). Application), which is incorporated herein by reference in its entirety.

In some embodiments, the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3, 2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile or a pharmaceutically acceptable salt thereof.

In some embodiments, the JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3, 2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile monohydrate.

((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-1-yl}tetrahydro-2H -pyran-2-yl)acetonitrile and characterization of its anhydride and monohydrate forms are described in US Patent Application No. 2014/0121198 (filed October 31, 2013) and US Patent Application No. 2015/ No. 0344497 (filed April 29, 2015), each of which is incorporated herein by reference in its entirety.

In some embodiments, the compounds of Table 1 are described in U.S. Patent Application No. 2011/0224190 (filed March 9, 2011), U.S. Patent Application No. 2014/0343030 (filed May 16, 2014), Patent Application No. 2014/0121198 (filed on October 31, 2013), US Patent Application No. 2010/0298334 (filed on May 21, 2010), US Patent Application No. 2011/0059951 (filed on August 31, 2010) ), US Patent Application No. 2012/0149681 (filed on November 18, 2011), US Patent Application No. 2012/0149682 (filed on November 18, 2011), US Patent Application No. 2013/0018034 (filed on November 18, 2012), 2013/0045963 (filed August 17, 2012) and US Patent Application No. 2014/0005166 (filed May 17, 2013). , each of these applications is incorporated herein by reference in its entirety.

In some embodiments, the JAK1 pathway inhibitor is described in US Patent Application No. 2011/0224190 (filed March 9, 2011), US Patent Application No. 2014/0343030 (filed May 16, 2014), Patent Application No. 2014/0121198 (filed on October 31, 2013), US Patent Application No. 2010/0298334 (filed on May 21, 2010), US Patent Application No. 2011/0059951 (filed on August 31, 2010) ), US Patent Application No. 2012/0149681 (filed on November 18, 2011), US Patent Application No. 2012/0149682 (filed on November 18, 2011), US Patent Application No. 2013/0018034 (filed on November 18, 2012), Compounds of US Patent Application No. 2013/0045963 (filed on August 17, 2012) and US Patent Application No. 2014/0005166 (filed on May 17, 2013) or pharmaceutically acceptable compounds thereof each of these applications is incorporated herein by reference in its entirety.

In some embodiments, the JAK1 pathway inhibitor is a compound of Formula I
or a pharmaceutically acceptable salt thereof, in which
X is N or CH,
L is C(=O) or C(=O)NH,
A is phenyl, pyridinyl, or pyrimidinyl, each optionally substituted with one or ^two independently selected R groups, and each ^R is independently fluoro or It is trifluoromethyl.

In some embodiments, the compound of Formula I is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[ 2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula I is 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazole-1 -yl]azetidin-1-yl}-N-[4-fluoro-2-(trifluoromethyl)phenyl]piperidine-1-carboxamide or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula I is [3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(1 -{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)azetidin-3-yl]acetonitrile or a pharmaceutically acceptable salt thereof.

In some embodiments, the JAK1 pathway inhibitor is a compound of Formula II
or a pharmaceutically acceptable salt thereof, in which
R ² is C _1-6 alkyl, C _1-6 haloalkyl, C _3-6 cycloalkyl, or C _3-6 cycloalkyl-C _1-3 alkyl, where the C _1-6 alkyl, C _3-6 cycloalkyl and C _3-6 cycloalkyl-C _1-3 alkyl are optionally substituted with 1, 2, or 3 substituents independently selected from fluoro, -CF ₃ and methyl;
R ³ is H or methyl;
R ⁴ is H, F or Cl;
^R5 is H or F;
^R6 is H or F;
^R7 is H or F;
R ⁸ is H or methyl;
R ⁹ is H or methyl;
R ¹⁰ is H or methyl, and R ¹¹ is H or methyl.

In some embodiments, the compound of Formula II is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl) azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof.

In some embodiments, the JAK1 pathway inhibitor is a compound of Formula III
or a pharmaceutically acceptable salt thereof, in which
Cy ⁴ is CN, OH, F, Cl, C _1-3 alkyl, C _1-3 haloalkyl, cyano-C _1-3 alkyl, HO-C _1-3 alkyl, amino, C _1-3 alkylamino and di- a tetrahydro-2H-pyran ring optionally substituted with one or two groups independently selected from (C _1-3 alkyl)amino, where the C _1-3 alkyl and di(C _{1-3 3} alkyl) amino is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, C _1-3 alkylaminosulfonyl and C _1-3 alkylsulfonyl;
R ¹² is -CH ₂ -OH, -CH(CH ₃ )-OH or -CH ₂ -NHSO ₂ CH ₃ .

In some embodiments, the compound of Formula III is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3, 2-b]pyridin-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile or a pharmaceutically acceptable salt thereof.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered in an amount of about 100 mg to about 600 mg per day on a free base basis. Accordingly, in some embodiments, the selective JAK1 pathway inhibitor is administered on a free base basis at about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, An amount of about 500 mg, about 550 mg or about 600 mg is administered.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered in an amount of about 200 mg per day on a free base basis.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered in an amount of about 300 mg per day on a free base basis.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered in an amount of about 400 mg per day on a free base basis.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered in an amount of about 500 mg per day on a free base basis.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered in an amount of about 600 mg per day on a free base basis.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered in an amount of about 200 mg on a free base basis once daily.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered in an amount of about 300 mg on a free base basis once daily.

In some embodiments, the JAK1 pathway inhibitor, or pharmaceutically acceptable salt thereof, is administered in an amount of about 400 mg once daily on a free base basis.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is administered in an amount of about 500 mg on a free base basis once daily.

In some embodiments, the JAK1 pathway inhibitor, or pharmaceutically acceptable salt thereof, is administered in an amount of about 600 mg once daily on a free base basis.

In some embodiments, the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is in one or more sustained release dosage forms, each containing the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof. administered in

Provided herein are methods of treating a subject cytokine-related disease or disorder in a subject in need of treatment, which method comprises administering a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof on a free base basis to wherein the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof comprises administering to the subject about 100 mg to 600 mg per day, wherein the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof comprises It is administered in one or more sustained release dosage forms.

The embodiments described herein are intended to be combined in any suitable combination, as if the embodiments were multiple dependent claims (e.g., selective JAK1 pathway inhibitors and dosages thereof). embodiments relating to any salt form of the compounds disclosed herein; embodiments relating to individual types of cytokine-related diseases or disorders; and embodiments relating to compositions and/or administration that may be combined in any combination). .

For example, as used herein, from the group consisting of cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell associated encephalopathy syndrome (CRES) in a subject. Provided are methods for treating selected cytokine-related diseases or disorders, the methods comprising administering to the subject {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4 -yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable thereof salt, administered once daily at a dose of about 200 mg on a free base basis, wherein the dose includes {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine -4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or its pharmaceutically acceptable One or more sustained release dosage forms are included, each containing a possible salt.

{1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d]pyrimidine) in sustained release dosage form -4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof (Table 1, Compound 1) is disclosed in US Patent No. 2015/0065484 (August 2014). No. 6, filed June 2006), which is incorporated herein by reference in its entirety.

All possible combinations are not shown separately here simply for the sake of brevity.

Compounds described herein can be asymmetric (eg, have one or more stereocenters). Unless otherwise indicated, all stereoisomers, including enantiomers and diastereomers, are intended. Compounds containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic form. Methods for preparing optically active forms from optically inactive starting materials are well known in the art, eg, by resolution of racemic mixtures or stereoselective synthesis. Many geometric isomers can also exist in the compounds described herein, such as olefins, C=N double bonds, and all such stable isomers are contemplated in the present invention. Cis and trans geometric isomers in the compounds of the invention are described and may be isolated as mixtures of isomers or as separated isomeric forms.

In some embodiments, the compound has the (R) configuration. In some embodiments, the compound has the (S) configuration.

Resolution of racemic mixtures of compounds can be performed by any of a number of methods well known in the art. Exemplary methods include fractional recrystallization using chiral splitting acids that are optically active salt-forming organic acids. Suitable resolving agents for the fractional recrystallization process are, for example, the D and L forms of various optically active camphorsulfonic acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or β-camphorsulfonic acid. It is an optically active acid such as. Other resolving agents suitable for fractional crystallization include stereomerically pure forms of α-methylbenzylamine (e.g. S and R forms or diastereomerically pure forms), 2-phenylglycinol , norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, and the like.

Resolution of racemic mixtures can also be carried out by elution on a column packed with an optically active resolving agent (eg dinitrobenzoylphenylglycine). Suitable elution solvent compositions can be determined by one of ordinary skill in the art.

The compounds described herein also include tautomeric forms. Tautomeric forms are obtained by exchange of adjacent double and single bonds and simultaneous proton transfer. Tautomeric forms include prototropic tautomers, which are protonated states of isomers with the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, as well as protons in which a proton can occupy two or more positions in a heterocyclic ring system. cyclic forms such as 1H-imidazole, 3H-imidazole, 1H-1,2,4-triazole, 2H-1,2,4-triazole and 4H-1,2,4-triazole, 1H-isoindole and 2H -isoindoles and 1H-pyrazoles and 2H-pyrazoles. Tautomeric forms may be in equilibrium or sterically fixed in one form by appropriate substitution.

The compounds described herein may also include isotopically labeled compounds of the present disclosure. An "isotopic" or "radiolabeled" compound is one in which one or more atoms have an atomic mass or mass number that is different from that normally found in nature (i.e., naturally occurring). or a compound of the present disclosure that is or is substituted by an atom having a mass number. Suitable radionuclides that may be incorporated into compounds of the present disclosure include ² H (also written with D as deuterium), ³ H (also written with T as tritium), ¹¹ C, ¹³ C, ¹⁴ C, Includes ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ¹⁸ F, ³⁵ S, ³⁶ Cl, ⁸² Br, ⁷⁵ Br, ⁷⁶ Br, ⁷⁷ Br, ¹²³ I, ¹²⁴ I, ¹²⁵ I and ¹³¹ I is not limited. For example, one or more hydrogen atoms in the compounds of the present disclosure can be replaced with a deuterium atom (e.g., C _1-6 of formula (I), (II), or (III) or the compounds of Table 1). One or more hydrogen atoms of the alkyl group can be optionally substituted with a deuterium atom, such that _-CD3 is substituted for _-CH3 ). As used herein, the term "compound" refers to all stereoisomers, geometric isomers, tautomers and isotopes of the depicted structure, unless the name indicates a specific stereoisomer. It means to include. Compounds herein identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms, unless specified otherwise.

All compounds and their pharmaceutically acceptable salts can be found together with other substances such as water and solvents (eg, hydrates and solvates) or can be isolated.

In some embodiments, the compounds described herein and their salts are substantially isolated. By "substantially isolated" is meant that the compound is at least partially or substantially separated from the context in which it was formed or detected. A partial separation can include, for example, a composition enriched in the compounds described herein. Substantial separation is at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, by weight of the compound or salt thereof described herein. It is possible to include compositions containing at least about 97% or at least about 99%. Methods for isolating compounds and their salts are common in the art.

As used herein, the term "pharmaceutically acceptable" means that the term "pharmaceutically acceptable" can be used in humans and animals without undue toxicity, irritation, allergic response or other problems or complications, within the scope of sound medical judgment. Refers to compounds, materials, compositions and/or dosage forms suitable for use in contact with tissue and commensurate with a reasonable benefit/risk ratio.

As used herein, the expressions "ambient temperature" and "room temperature" or "rt" are understood in the art and generally refer to the temperature of the room in which the reaction is conducted, e.g., from about 20°C to about 30°C. , for example, the reaction temperature.

The present invention also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, "pharmaceutically acceptable salts" are derivatives of the disclosed compounds in which the parent compound is modified by converting an existing acid or base moiety to its salt form. has been done. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic salts of basic residues such as amines, alkaline or organic salts of acidic residues such as carboxylic acids, etc. . Pharmaceutically acceptable salts of the invention include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts of the invention can be synthesized from a parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of the two. It can be prepared by Generally, non-aqueous solvents such as ether, ethyl acetate, alcohol (eg methanol, ethanol, isopropanol, or butanol) or acetonitrile (ACN) are preferred. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed. , Mack Publishing Company, Easton, Pa. , 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

As used herein, the terms "subject," "individual," or "patient," used interchangeably, refer to mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, Refers to any animal including a pig, cow, sheep, horse or primate, most preferably a human. In some embodiments, a "subject," "individual," or "patient" is in need of a treatment as described above.

In some embodiments, the inhibitor is administered in a therapeutically effective amount. As used herein, the expression "therapeutically effective amount" refers to the biological response or response sought in a tissue, system, animal, individual, or human by a researcher, veterinarian, physician, or other clinician. Refers to the amount of active compound or drug that elicits a drug response. In some embodiments, the dosage of the compound or pharmaceutically acceptable salt thereof administered to a patient or individual is about 1 mg to about 2 g, about 1 mg to about 1000 mg, about 1 mg to about 500 mg, about 1 mg to about about 200 mg, about 1 mg to about 100 mg, about 1 mg to 50 mg, or about 50 mg to about 500 mg. In some embodiments, the dosage of the compound or pharmaceutically acceptable salt thereof is about 200 mg.

As used herein, the term "treat" or "treatment" refers to (1) inhibiting a disease, e.g., in an individual experiencing or exhibiting the pathology or symptoms of a disease, condition, or disorder; (2) ameliorating a disease, such as reducing the severity of a disease, symptom or disorder; ameliorating the disease, condition, or disorder (i.e., halting the progression of the condition and/or symptom) in an individual experiencing or exhibiting the condition or symptom of the disorder; or (3) predisposing to the disease, condition, or disorder. refers to one or more of the prevention of a disease, condition, or disorder in an individual who is not yet experiencing or exhibiting the pathology or symptoms of that disease. In some embodiments, treatment refers to inhibiting or ameliorating the disease. In some embodiments, treating is preventing disease.

Combination Therapy The methods described herein can further include administering one or more additional therapeutic agents. One or more additional therapeutic agents can be administered to the patient simultaneously or sequentially.

In some embodiments, the additional therapeutic agent is an IL-6 antagonist or receptor antagonist. In some embodiments, the IL-6 receptor antagonist is tocilizumab.

In some embodiments, the additional therapeutic agent is an inhibitor of MCP-1. In some embodiments, the additional therapeutic agent is an inhibitor of MIP1B. In some embodiments, the additional therapeutic agent is an inhibitor of IL-2R. In some embodiments, the additional therapeutic agent is an inhibitor of IL-1R. In some embodiments, the additional therapeutic agent is an inhibitor of TNF-α.

In some embodiments, the additional therapeutic agent is an anti-CD25 antibody. In some embodiments, the anti-CD25 antibody is daclizumab.

In some embodiments, the additional therapeutic agent is an antagonist of IL-1β.

In some embodiments, the additional therapeutic agent is an IL1 receptor antagonist (IL1Ra). In some embodiments, the IL1 receptor antagonist (IL1Ra) is anakinra.

In some embodiments, the additional therapeutic agent is a corticosteroid. In some embodiments, the corticosteroid is prednisone.

In some embodiments, any of the aforementioned additional therapeutic agents are used in further combination with a corticosteroid (eg, prednisone).

In some embodiments, additional therapeutic agents include tocilizumab and corticosteroids. In some embodiments, the additional therapeutic agents include tocilizumab and prednisone.

Pharmaceutical Formulations and Dosage Forms When used as a pharmaceutical, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof can be administered in the form of a pharmaceutical composition. These compositions can be prepared by methods well known in the pharmaceutical art and can be administered by a variety of routes, depending on whether local or systemic treatment is desired and the area to be treated. be. Administration can be topical (transdermal, epithelial, ophthalmic and mucosal, including intranasal, vaginal and rectal delivery), pulmonary (e.g. by inhalation or insufflation of a powder or aerosol, such as through a nebulizer; can be intraluminal or intranasal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular or injection, or infusion, or intracranial, such as intrathecally or intraventricularly. Parenteral administration can be in the form of a single bolus dose or by, for example, a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous bases, powder bases, oil bases or thickening agents, and the like may be necessary or desirable.

The present invention includes a JAK1 pathway inhibitor as described herein or a pharmaceutically acceptable drug thereof as an active ingredient in combination with one or more pharmaceutically acceptable carriers (excipients). Also included are pharmaceutical compositions containing salts. In some embodiments, the composition is suitable for topical administration. In making this composition, the active ingredient is typically mixed with an excipient, diluted by the excipient, or placed in such form, for example, in the form of a capsule, sachet, paper or other container. Enclosed in a carrier. When the excipient functions as a diluent, it can be a solid, semi-solid or liquid material that acts as a solvent, carrier or medium for the active ingredient. The composition may therefore be used as a tablet, pill, powder, troche, sachet, cachet, elixir, suspension, emulsion, solution, syrup, aerosol (as a solid medium or in a liquid medium), e.g. They may be in the form of ointments, soft gelatin capsules, hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders containing % by weight of the active compound.

In preparing the formulation, the active compound can be milled to provide the appropriate particle size before combining with the other ingredients. If the active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh. If the active compound is substantially water soluble, the particle size can be adjusted by milling to, for example, about 40 mesh to provide substantially uniform distribution in the formulation.

The JAK1 pathway inhibitor can be milled using known milling procedures such as wet milling to obtain appropriate particle sizes for tablet formation and for other formulation types. Finely divided (nanoparticle) preparations of JAK1 selective inhibitors can be prepared by methods well known in the art, see for example International Patent Application No. WO 2002/000196. thing.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, Water, syrup and methylcellulose are mentioned. Additionally, the formulations may contain lubricating agents, wetting agents, emulsifying and suspending agents such as talc, magnesium stearate and mineral oil, preservatives such as methyl benzoate and propyl hydroxybenzoate, sweetening and flavoring agents. The compositions of the invention can be formulated to provide rapid, sustained, or delayed release of the active ingredient after administration to a patient by using methods known in the art.

The compositions may be formulated in unit dosage form, with each dose containing about 5 to about 1000 mg (1 g), more usually about 100 to about 500 mg of active ingredient. The term "unit dosage form" refers to physically discrete units suitable as unit dosages for human subjects and other mammals, each unit being associated with suitable pharmaceutical excipients. , a predetermined amount of active substance calculated to produce the desired therapeutic effect.

In some embodiments, compositions of the invention contain from about 5 to about 50 mg of active ingredient. For those skilled in the art, this means about 5 to about 10, about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about It will be appreciated that embodiments of the composition include 40 to about 45 or about 45 to about 50 mg of active ingredient.

In some embodiments, compositions of the invention contain about 50 to about 500 mg of active ingredient. For those skilled in the art, this means about 50 to about 100, about 100 to about 150, about 150 to about 200, about 200 to about 250, about 250 to about 300, about 350 to about 400 or about 450 to about 500 mg of activity. It will be understood that it is an embodiment of a composition that includes the ingredients.

In some embodiments, compositions of the invention contain about 500 to about 1000 mg of active ingredient. For those skilled in the art, this means about 500 to about 550, about 550 to about 600, about 600 to about 650, about 650 to about 700, about 700 to about 750, about 750 to about 800, about 800 to about 850, about It will be appreciated that embodiments of the compositions include from 850 to about 900, from about 900 to about 950, or from about 950 to about 1000 mg of active ingredient.

Similar dosages of the compounds described herein can be used in the methods and uses of the invention.

The active compounds can be effective over a wide dosage range and are therefore generally administered in pharmaceutically effective amounts. However, the actual amount of compound administered will usually depend on the condition being treated, the route of administration chosen, the actual compound administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc. It will be understood that this will be determined by the physician according to the relevant circumstances.

For preparing solid compositions such as tablets, the primary active ingredient is mixed with pharmaceutical excipients to form a solid preformulated composition containing a homogeneous mixture of the compounds of the invention. When these preformulated compositions are referred to as homogeneous, the active ingredient is typically uniformly dispersed throughout the composition, so that the composition can be divided into equally effective tablets, pills, and capsules. It can be easily subdivided into unit dosage forms. This solid preformulation is further subdivided into unit dosage forms of the type described above containing, for example, from about 0.1 to about 1000 mg of the active ingredient of the invention.

The tablets or pills of the invention can be coated or otherwise mixed to provide a dosage form that provides the advantage of prolonging action. For example, the tablet or pill can include an inner dosage component and an outer dosage component, the outer component being in the form of an envelope relative to the inner component. These two components can be separated by an enteric layer. This enteric layer functions to prevent disintegration in the stomach and also allows the inner components to reach the duodenum intact or for delayed release. A variety of materials can be used for such enteric layers or coatings, including many polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol and cellulose acetate. .

Liquid forms that may be incorporated for oral or injectable administration of the compounds and compositions of the invention include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and cottonseed oil, sesame oil, coconut oil or peanut oil. Includes emulsions flavored with edible oils such as oils, elixirs and similar pharmaceutical solvents.

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions may include suitable pharmaceutically acceptable excipients as described above. In some embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. The composition can be nebulized using an inert gas. Nebulized solutions can be inhaled directly from the nebulizing device, or the nebulizing device can be attached to a face mask, obturator, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered orally or nasally from devices that deliver the formulation in a suitable manner.

Topical formulations may include one or more conventional carriers. In some embodiments, ointments can include water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ethers, propylene glycol, white petrolatum, and the like. The carrier composition of the cream may be based on a combination of water, glycerol, and one or more other ingredients such as, for example, glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. Gels may be formulated using isopropyl alcohol and water, preferably in combination with other components such as glycerol, hydroxyethyl cellulose, etc. In some embodiments, the topical formulation contains at least about 0.1, at least about 0.25, at least about 0.5, at least about 1, at least about 2 or at least about 5% by weight of a compound described herein. Contains compounds that Topical formulations may be conveniently packaged in 100 g tubes, optionally with instructions for treating selected indications, such as, for example, psoriasis or other skin conditions.

The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of administration, such as prophylaxis or therapy, the condition of the patient, the method of administration, and the like. In therapeutic applications, the compositions may be administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially suppress the symptoms of the disease and its complications. Effective doses depend on the symptoms of the disease being treated and the judgment of the attending clinician based on factors such as the severity of the disease, the age, weight, and general condition of the patient.

The composition administered to a patient may be in the form of a pharmaceutical composition as described above. These compositions may be sterilized by conventional sterilization techniques or may be sterile filtered. Although aqueous solutions can be packaged or lyophilized for use directly, lyophilized preparations are combined with a sterile aqueous carrier prior to administration. The pH of the compound preparation will typically be between 3 and 11, more preferably between 5 and 9, and most preferably between 7 and 8. It will be appreciated that the use of certain of the aforementioned excipients, carriers or stabilizers results in formulations of pharmaceutical salts.

The therapeutic amount of a compound of the invention may vary depending on, for example, the particular application being treated, the method of administering the compound, the health and condition of the patient and the judgment of the prescribing physician. The proportions or concentrations of compounds described herein in pharmaceutical compositions can vary depending on many factors, including dosage, chemical nature (eg, hydrophobicity), and route of administration. For example, the compounds described herein can be provided in an aqueous physiological buffer solution containing about 0.1 to about 10% w/v of the compound for parenteral administration. Some typical dosage ranges are from about 1 μg/kg to about 1 g/kg of body weight per day. In some embodiments, the dosage range is from about 0.01 mg/kg to about 100 mg/kg of body weight per day. Dosage will depend on variables such as the type and extent of the disease or disorder, the overall health of the particular patient, the relative biological effectiveness of the selected compound, the formulation of excipients and its route of administration. may depend on. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

The compositions of the invention can further include one or more additional pharmaceutical agents such as chemotherapeutic agents, steroids, anti-inflammatory compounds or immunosuppressants, examples of which are included herein. ing.

Kits The invention also includes pharmaceutical kits useful in treating and/or preventing cytokine-related diseases or disorders, such as, for example, CRS, which kits contain the therapeutically effective agents described herein. Also included are one or more containers containing a pharmaceutical composition containing an amount of the compound. Such kits optionally include one or more of a variety of conventional pharmaceutical kit components, such as, for example, containers containing one or more pharmaceutically acceptable carriers, additional containers, etc. Further may be included and will be readily apparent to those skilled in the art. Instructions, either as an insert or as a label, indicating amounts of such components, guidelines for administration and/or guidelines for mixing the components may also be included in the kit.

The invention will be described in more detail by means of specific examples. The following examples are provided for illustrative purposes and are not intended to limit the invention in any way. Those skilled in the art will readily recognize various non-critical parameters that can be changed or modified to produce essentially the same result. The compounds of the Examples have been found to be JAK inhibitors by at least one assay described herein.

Example A: In vitro JAK Kinase Assay JAK1 pathway inhibitors that can be used to treat cytokine-related diseases or disorders were described by Park et al. JAK targets are tested for inhibitory activity according to the following in vitro assay described in , Analytical Biochemistry 1999, 269, 94-104. The catalytic domains of human JAK1 (a.a. 837-1142), JAK2 (a.a. 828-1132) and JAK3 (a.a. 781-1124) with an N-terminal His tag were baculotransduced in insect cells. It is expressed using a virus and purified. The catalytic activity of JAK1, JAK2 or JAK3 was assayed by measuring the phosphorylation of biotinylated peptides. Phosphorylated peptides were detected by homogeneous time-resolved fluorescence (HTRF). In a 40 μL reaction containing enzyme, ATP, and 500 nM peptide in 50 mM Tris (pH 7.8) buffer containing 100 mM NaCl, 5 mM DTT, and 0.1 mg/mL (0.01%) BSA. The IC ₅₀ of the compound is determined for each kinase. For an IC ₅₀ measurement of 1 mM, the ATP concentration in the reaction is 1 mM. Reactions were run for 1 hour at room temperature and then stopped with 20 μL of 45 mM EDTA, 300 nM SA-APC, 6 nM Eu-Py20 assay buffer (Perkin Elmer, Boston, MA). Binding to europium-labeled antibodies was performed for 40 minutes and HTRF signals were measured on a Fusion plate reader (Perkin Elmer, Boston, MA). The compounds in Table 1 were tested in this assay and were shown to have the IC ₅₀ values seen in Table 1.

Example B: Anti-CD3 Antibody-Induced Cytokine Release Syndrome in Balb/c Mice JAK1 pathway inhibitors are described by Ferran, C.; et al. Clin. Exp. Immunol. It is possible to test for efficacy against CRS according to the in vivo assay described in 1991, 86, 537-543. Specifically, this study allows testing the ability of compounds to reduce or ameliorate anti-CD3 antibody-induced cytokine release syndrome (CRS) in BALB/c mice. The antibody, clone 145-2C11, is an immunoglobin G (IgG) hamster MoAb specific for the epsilon chain of the CD3 mouse molecule (Leo, O. et al., Proc. Natl. Acad. Sci. USA, 1987, 34 , 1374). Treatment with 145-2C11 induces high-affinity IL-2 receptors on the surface of splenic T cells and induces tumor necrosis factor (TNF-α), IL-2, IL-3, IL-6 and interferon-gamma ( IFN-γ) (Ferran, et al. Eur. J. Immunol. 1990, 20, 509-515, and Algre, M. et al., Eur. J. Immunol., 1990, 707). Release of these cytokines results in behavioral changes in the animal (eg, inactivity, piloerection, etc.).

A. Materials and methods
^A Compound 1 of Table 1 or {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d] Synthesis and preparation of pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile and its adipate salt are disclosed in U.S. Patent Application No. 2011/0224190 (filed March 9, 2011). , can be found in U.S. Patent Application No. 2013/0060026 (filed September 6, 2012) and U.S. Patent Application No. 2014/0256941 (filed March 5, 2014), each of which is incorporated by reference in its entirety. incorporated herein by.

B. Experimental Design The primary objective of this study was to test the ability of JAK1 pathway inhibitors (e.g., Compound 1) to reduce or ameliorate anti-CD3 antibody-induced cytokine release syndrome (CRS) in BALB/c mice. Ta. A total of 32 BALB/c mice were used in this one-day study. Animals were weighed and monitored during the experiment prior to administering test material. On day 0, 1 hour before anti-CD3 antibody administration, vehicle (0.5% methylcellulose) or Compound 1 was administered by oral gavage (PO) to animals in groups 2-4 as detailed in Table 1A. A single dose was administered. Group 1 served as an untreated control and received no treatment. One hour after pretreatment with vehicle or Compound 1, animals in groups 2-4 received 10 μg of anti-CD3ε antibody (clone 145-2C11) by intravenous injection (IV) to induce CRS. All animals were euthanized by _CO2 inhalation 1.5 hours after anti-CD3 administration. Whole blood was collected by cardiac puncture into K ₂ EDTA blood collection tubes and plasma was stored on ice until processing. Plasma was collected and stored at -80°C until cytokine multiplexing.

C. Experimental Procedure I Test Article Pretreatment On day 0, animals were administered vehicle or test article or Compound 1 as shown in Table 1A. Group 2 received a single dose of vehicle (0.5% methylcellulose) via PO at 0.1 mL/20 g. Group 3 received a single dose of 60 mg/kg Compound 1 PO at 0.1 mL/20 g. Group 4 received a single dose of Compound 1 at 120 mg/kg PO in 0.1 mL/20 g. Group 1 served as an untreated control and received no treatment.

II Administration of anti-CD3ε antibody One hour after test substance administration, anti-CD3ε antibody (clone 145-2C11) was administered to groups 2-4 by intravenous injection. Each animal in groups 2-4 received 10 μg of anti-CD3ε antibody in 0.1 mL.

III. Survival Monitoring After administration of anti-CD3 antibodies, animals were carefully monitored for signs of distress due to the developed systemic inflammatory response. Animals that failed to rise, were cold to the touch, or were moribund were euthanized. Moribund animals were euthanized by _CO2 inhalation, blood was collected by cardiac puncture, and plasma was saved.

IV Sacrifice One and a half hours after anti-CD3 antibody administration, all animals were euthanized by _CO2 inhalation.

V Sample Collection At sacrifice, whole blood was collected from each animal by cardiac puncture into _K2EDTA blood collection tubes. Blood was centrifuged and plasma was collected into cryovials. Plasma was frozen and stored at -80°C for downstream cytokine multiplex assays.

VI Cytokine multiplex analysis Plasma samples are thawed on ice and used for cytokine multiplexing according to the manufacturer's protocol (ThermoFisher).

D Results Compound 1 inhibited IL-6 concentrations in the blood compartment in a dose-dependent manner (Figure 1). This serves as confirmation of the biological activity observed in the preclinical model of Con A described in Example C below. One-way analysis of variance (unpaired) (ANOVA) incorporating Sidak's multiple comparison test was performed using GraphPad Prism (version 4.00; GraphPad Software, San Diego California, USA). Values of p<0.05 were considered significant.

Example C: Concanavalin A-Induced Cytokine Release Syndrome Concanavalin A (Con A) is a selective T lymphocyte mitogen that results in the release of a wide range of inflammatory cytokines and proliferation of CD4 and CD8 T cells. Injection of Con-A has been shown in the literature to induce cytokine release syndrome and serves as a model to use in testing the efficacy of treatments for cytokine release syndrome (Gantner, F. at al. Hepatology, 1995 , 21, 190-198). Mitogenic responses are dependent on T cell receptor expression. Animals exhibit behavioral changes such as fever, malaise, hypotension, hypoxia, capillary leak and potential multiorgan toxicity.

A. Materials and methods

B Experimental Design Specifically, this study tests the ability of selective JAK1 inhibitors (e.g., Compound 1, Table 1) to reduce or ameliorate Con A-induced cytokine release syndrome (CRS) in BALB/c mice. A total of 40 BALB/c mice were used in this one-day study. Animals were weighed and monitored during the experiment prior to administering test material. On day 0, 60 minutes before Con A administration, vehicle (0.5% methylcellulose) or Compound 1 (60 and 120 mg/kg) was gavaged to animals in groups 2-4 as detailed in Table 2A. Administration (PO), a single dose was administered. Group 1 served as an untreated control and received no treatment. Forty-five minutes after pretreatment with vehicle or Compound 1, animals in groups 2-4 received 20 mg/kg Con A by intravenous injection (IV) to induce CRS. Two hours after administering Con A, all animals were euthanized by _CO2 inhalation. Whole blood was collected by cardiac puncture into K ₂ EDTA blood collection tubes and plasma was stored on ice until processing. Plasma was collected and stored at -80°C until cytokine multiplexing.

C. Experimental Procedures Day 1 Animals were weighed and the dose of Con-A (20 mg/kg) was calculated.
Solvent and Compound 1 were prepared in corresponding doses.

Day 0 I Test Article Pretreatment On day 0, animals were administered vehicle or Compound 1 as shown in Table 2A. Group 1 served as an untreated control and received no treatment. Group 2 received a single dose of vehicle (0.5% methylcellulose) via PO at 0.1 mL/20 g. Group 3 received a single dose of 60 mg/kg Compound 1 PO at 0.1 mL/20 g. Group 4 received a single dose of Compound 1 at 120 mg/kg PO in 0.1 mL/20 g.

II Administration of Con-A Sixty minutes after test substance administration, Con-A was administered to groups 2-4 by intravenous injection. Each animal in groups 2-4 received 0.2 mL of Con-A 20 mg/kg.

III. Survival Monitoring After administration of Con-A, animals were carefully monitored for signs of distress due to the developed systemic inflammatory response.

IV Sacrifice Two hours after Con-A administration, all animals were euthanized by CO ₂ inhalation.

V Sample Collection At sacrifice, whole blood was collected from each animal by cardiac puncture into _K2EDTA blood collection tubes. Blood was centrifuged and plasma was collected into cryovials. Plasma was frozen and stored at -80°C for downstream cytokine multiplex assays.

VI Cytokine multiplex analysis Plasma samples are thawed on ice and used for cytokine multiplexing according to the manufacturer's protocol (ThermoFisher).

D Results Compound 1 inhibited IL-6 concentrations in the blood compartment in a dose-dependent manner (FIG. 2A). This cytokine is key related to the pathophysiology of CRS. T cell-derived IFNγ and GM-CSF cytokines were also significantly inhibited, suggesting that compound 1 has therapeutic potential beyond the mechanism of action limited by tocilizumab (anti-IL-6R only) (Figures 2B and 2C).

Monocyte- and/or macrophage-derived cytokines were also reduced. A statistically significant dose-dependent decrease in IL-12 (Figure 3A) and a trend towards treatment effects with IL-1β (Figure 3B) and IL-18 (Figure 3C) were observed, indicating that JAK1-specific inhibition significantly reduced CRS. It was suggested that this drug has therapeutic potential that goes beyond the immune cell types involved in pathology.

Importantly, the cytokine IL-5 (Figure 4) is unaffected by Compound 1 treatment, is independent of JAK1, and is not involved in CRS pathology. This data suggests that efficacy with Compound 1 is not mediated by broad, nonspecific immunosuppression.

One-way analysis of variance (unpaired) (ANOVA) incorporating Sidak's multiple comparison test was performed using GraphPad Prism (version 4.00, GraphPad Software, San Diego California, USA). Values of p<0.05 were considered significant.

Example D: Preparation of sustained release formulation of Compound 1 Sustained release tablets containing Compound 1 were prepared with excipients in the amounts shown in the table below. Protocol A was used for SR1 tablets, protocol B was used for SR2 tablets, protocol C was used for SR3 tablets and 25 mg SR tablets, and protocol D was used for SR4 tablets. These procedures are disclosed in US Patent Application No. 2015/0065484, which relates to sustained release dosage forms of Compound 1.

Protocol A
Step 1 Separately sieve the adipate salt of Compound 1, microcrystalline cellulose, hypromellose (Methocel K100 LV and Methocel K4M) and lactose monohydrate.
Step 2 Transfer the sifted ingredients from Step 1 to a suitable blender and mix.
Step 3 Transfer the mixture from Step 2 to a suitable granulator and mix.
Step 4 Add purified water while mixing.
Step 5 Transfer the granules from Step 4 to a suitable dryer and dry until the LOD is less than 3%.
Step 6 Sift the granules from Step 5.
Step 7 Mix the granules from Step 6 with the sieved magnesium stearate in a suitable blender.
Step 8 Compress the final mixture of Step 7 in a suitable rotary tablet machine.

Protocol B
Step 1 Separately sieve the adipate salt of Compound 1, microcrystalline cellulose, hypromellose, and pregelatinized starch.
Step 2 Transfer the sifted ingredients from Step 1 to a suitable blender and mix.
Step 3 Transfer the mixture from Step 2 to a suitable granulator and mix.
Step 4 Add purified water while mixing.
Step 5 Transfer the granules from Step 4 to a suitable dryer and dry until the LOD is less than 3%.
Step 6 Sift the granules from Step 5.
Step 7 Separately sieve the polyox, butylated hydroxytoluene and colloidal silicon dioxide.
Step 8 Transfer the granules from Step 6 and the ingredients from Step 7 to a suitable blender and mix.
Step 9 Add the sifted magnesium stearate to the Step 8 ingredients and continue blending.
Step 10 Compress the final mixture of Step 9 on a suitable rotary tablet machine.

Protocol C
Step 1 Separately sieve lactose monohydrate, adipate salt of Compound 1, microcrystalline cellulose and hypromellose through a suitable sieve.
Step 2 Transfer the sifted ingredients from Step 1 to a suitable blender and mix.
Step 3 Transfer the mixture from Step 2 to a suitable granulator and mix.
Step 4 Add purified water while mixing.
Step 5 Sift the wet granules through a suitable sieve.
Step 6 Transfer the granules from Step 5 to a suitable dryer and dry until the LOD is less than 3%.
Step 7 Mill the granules from Step 6.
Step 8 Mix the granules from Step 7 with the sieved magnesium stearate in a suitable blender.
Step 9 Compress the final mixture of Step 8 on a suitable rotary tablet machine.

Protocol D
Step 1 Separately sieve the pregelatinized starch, adipate salt of Compound 1, hypromellose and a portion of the required microcrystalline cellulose through a suitable sieve.
Step 2 Transfer the sifted ingredients from Step 1 to a suitable blender and mix.
Step 3 Transfer the mixture from Step 2 to a suitable granulator and mix.
Step 4 Add purified water while mixing.
Step 5 Sift the wet granules through a suitable sieve.
Step 6 Transfer the granules from Step 5 to a suitable dryer and dry until the LOD is less than 3%.
Step 7 Mill the granules from Step 6.
Step 8 Sift the remaining portion of microcrystalline cellulose and half of the sodium bicarbonate.
Step 9 Transfer the milled granules from Step 7 and the ingredients from Step 8 to a suitable blender and mix.
Step 10 Sift the remaining portion of the sodium bicarbonate and mix with the mixture of Step 9.
Step 11 Sift the magnesium stearate and mix with the mixture of Step 10.
Step 12 Compress the final mixture of Step 11 on a suitable rotary tablet machine.

SR1: Composition of 100mg sustained release tablet
The conversion factor for ^a adipate to free base is 0.7911.
^b Added after granulation.
^c was removed during treatment.

SR2: Composition of 100mg sustained release tablet
The conversion factor for ^a adipate to free base is 0.7911.
^b Added after granulation.
^c was removed during treatment.

SR3 (100mg): Composition of 100mg sustained release tablet
The conversion factor for ^a adipate to free base is 0.7911.
^b Added after granulation.
^c was removed during treatment.

SR4: Composition of 100mg sustained release tablet
The conversion factor for ^a adipate to free base is 0.7911.
^b Added after granulation.
^c was removed during treatment.
^d A portion was added before granulation, and a portion was added after granulation.

25mg SR: Composition of 25mg sustained release tablet
The conversion factor for ^a adipate to free base is 0.7911.
^b Added after granulation.
^c was removed during treatment.

Various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to come within the scope of the appended claims. Each reference, including all patents, patent applications, and publications cited in this application is incorporated by reference in its entirety.

Various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to come within the scope of the appended claims. Each reference, including all patents, patent applications, and publications cited in this application is incorporated by reference in its entirety.
Further, the present application includes the following aspects.
[Aspect 1]
A method of treating a cytokine-related disease or disorder in a subject, said method comprising administering to said subject a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof.
[Aspect 2]
2. The method of embodiment 1, wherein the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is selective for JAK1 over JAK2, JAK3, and Tyk2.
[Aspect 3]
Aspect 1, wherein the cytokine-related disease or disorder is cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell-related encephalopathy syndrome (CRES). Or the method described in 2.
[Aspect 4]
4. The method according to aspect 3, wherein the cytokine-related disease or disorder is cytokine release syndrome (CRS).
[Aspect 5]
4. The method according to aspect 3, wherein the cytokine-related disease or disorder is hemophagocytic lymphohistiocytosis (HLH).
[Aspect 6]
4. The method according to aspect 3, wherein the cytokine-related disease or disorder is macrophage activation syndrome (MAS).
[Aspect 7]
7. The method of aspect 6, wherein the macrophage activation syndrome (MAS) is associated with systemic juvenile idiopathic arthritis.
[Aspect 8]
4. The method of aspect 3, wherein the cytokine-related disease or disorder is CAR-T cell-related encephalopathy syndrome (CRES).
[Aspect 9]
The JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d] 9. The method according to any one of aspects 1 to 8, wherein the method is pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof.
[Aspect 10]
The JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d] 9. The method according to any one of aspects 1 to 8, wherein the method is pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate.
[Aspect 11]
The JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]- According to any one of embodiments 1 to 8, which is 2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof. Method described.
[Aspect 12]
The JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]- The method according to any one of aspects 1 to 8, wherein the method is 2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide phosphate.
[Aspect 13]
The JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1 -yl}tetrahydro-2H-pyran-2-yl)acetonitrile or a pharmaceutically acceptable salt thereof.
[Aspect 14]
The JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1 -yl}tetrahydro-2H-pyran-2-yl)acetonitrile monohydrate.
[Aspect 15]
15. The method according to any one of aspects 1-14, further comprising administering tocilizumab to said subject.
[Aspect 16]
15. The method according to any one of aspects 1-14, further comprising administering a corticosteroid to said subject.
[Aspect 17]
15. The method according to any one of aspects 1-14, further comprising administering prednisone to said subject.
[Aspect 18]
15. The method according to any one of aspects 1-14, further comprising administering tocilizumab and a corticosteroid to said subject.

Claims (18)

A method of treating a cytokine-related disease or disorder in a subject, said method comprising administering to said subject a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof. 2. The method of claim 1, wherein the JAK1 pathway inhibitor or pharmaceutically acceptable salt thereof is selective for JAK1 over JAK2, JAK3, and Tyk2. 12. The cytokine-related disease or disorder is cytokine release syndrome (CRS), hemophagocytic lymphohistiocytosis (HLH), macrophage activation syndrome (MAS), or CAR-T cell associated encephalopathy syndrome (CRES). The method described in 1 or 2. 4. The method of claim 3, wherein the cytokine-related disease or disorder is cytokine release syndrome (CRS). 4. The method of claim 3, wherein the cytokine-related disease or disorder is hemophagocytic lymphohistiocytosis (HLH). 4. The method of claim 3, wherein the cytokine-related disease or disorder is macrophage activation syndrome (MAS). 7. The method of claim 6, wherein the macrophage activation syndrome (MAS) is associated with systemic juvenile idiopathic arthritis. 4. The method of claim 3, wherein the cytokine-related disease or disorder is CAR-T cell associated encephalopathy syndrome (CRES). The JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d] The method according to any one of claims 1 to 8, wherein the method is pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile or a pharmaceutically acceptable salt thereof. The JAK1 pathway inhibitor is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidin-4-yl}-3-[4-(7H-pyrrolo[2,3-d] The method according to any one of claims 1 to 8, which is pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-3-yl}acetonitrile adipate. The JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]- Any one of claims 1 to 8, which is 2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof. The method described in. The JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]- The method according to any one of claims 1 to 8, which is 2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide phosphate. The JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1 -yl}tetrahydro-2H-pyran-2-yl)acetonitrile or a pharmaceutically acceptable salt thereof. The JAK1 pathway inhibitor is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1 -yl}tetrahydro-2H-pyran-2-yl)acetonitrile monohydrate. 15. The method of any one of claims 1-14, further comprising administering tocilizumab to the subject. 15. The method of any one of claims 1-14, further comprising administering a corticosteroid to the subject. 15. The method of any one of claims 1-14, further comprising administering prednisone to the subject. 15. The method of any one of claims 1-14, further comprising administering tocilizumab and a corticosteroid to the subject.